U.S. patent number 10,314,941 [Application Number 15/895,518] was granted by the patent office on 2019-06-11 for preventative therapy for post-traumatic osteoarthritis.
This patent grant is currently assigned to University of Iowa Research Foundation. The grantee listed for this patent is University of Iowa Research Foundation. Invention is credited to Marc Brouillette, Mitchell Coleman, Tae-Hong Lim, James A. Martin, Todd O. McKinley.
United States Patent |
10,314,941 |
McKinley , et al. |
June 11, 2019 |
Preventative therapy for post-traumatic osteoarthritis
Abstract
Compositions comprising a reverse-temperature sensitive hydrogel
comprising a biopolymer such as a polysaccharide and a synthetic
polymer, and a compound in an amount that reversibly inhibits
respiratory enzyme complex I, and methods of using the composition,
are provided.
Inventors: |
McKinley; Todd O.
(Indianapolis, IN), Martin; James A. (Iowa City, IA),
Coleman; Mitchell (Iowa City, IA), Lim; Tae-Hong
(Coralville, IA), Brouillette; Marc (Coralville, IA) |
Applicant: |
Name |
City |
State |
Country |
Type |
University of Iowa Research Foundation |
Iowa City |
IA |
US |
|
|
Assignee: |
University of Iowa Research
Foundation (Iowa City, IA)
|
Family
ID: |
56801838 |
Appl.
No.: |
15/895,518 |
Filed: |
February 13, 2018 |
Prior Publication Data
|
|
|
|
Document
Identifier |
Publication Date |
|
US 20180169298 A1 |
Jun 21, 2018 |
|
Related U.S. Patent Documents
|
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
PCT/US2016/047360 |
Aug 17, 2016 |
|
|
|
|
62207059 |
Aug 19, 2015 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
47/36 (20130101); A61K 47/38 (20130101); A61K
9/0024 (20130101); A61L 27/50 (20130101); A61K
47/34 (20130101); A61K 31/515 (20130101); A61L
27/54 (20130101); A61P 19/02 (20180101); A61L
27/26 (20130101); A61K 31/155 (20130101); A61L
27/52 (20130101); A61K 9/06 (20130101); A61L
27/26 (20130101); C08L 5/08 (20130101); A61L
27/26 (20130101); C08L 71/02 (20130101); A61L
2300/434 (20130101); A61L 2430/24 (20130101); A61L
2400/06 (20130101) |
Current International
Class: |
A61K
9/00 (20060101); C08L 71/02 (20060101); A61K
31/155 (20060101); A61K 31/515 (20060101); A61P
19/02 (20060101); A61K 47/34 (20170101); C08L
5/08 (20060101); A61K 9/06 (20060101); A61L
27/54 (20060101); A61L 27/52 (20060101); A61L
27/50 (20060101); A61L 27/26 (20060101); A61K
47/36 (20060101); A61K 47/38 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
|
|
|
|
|
|
|
108136070 |
|
Jun 2018 |
|
CN |
|
1496037 |
|
Jan 2005 |
|
EP |
|
WO-2006047279 |
|
May 2006 |
|
WO |
|
WO-2010061005 |
|
Jun 2010 |
|
WO |
|
WO-2013171736 |
|
Nov 2013 |
|
WO |
|
WO-2017031214 |
|
Feb 2017 |
|
WO |
|
Other References
Tsuyoshi (EP1496037 A1) (Year: 2005). cited by examiner .
Mckinley et al. ("Mitochondrial Based Treatments that Prevent
Post-Traumatic Osteoarthritis in a Translational Large Animal
Intraarticular Fracture Survival Model", 2013). (Year: 2013). cited
by examiner .
Martin, James, "Blocking Acute Oxidative Insult to Chondrocytes
Prevents Post-Traumatic Osteoarthritis in a Porcine Model of Tibial
Plafond Fracture (Abstract of Presentation)", Extremity and War
Injuries XI Conference, Washington DC, (2016), 1pg. cited by
applicant .
"International Application Serial No. PCT/US2016/047360,
International Search Report dated Nov. 21, 2016", 4 pgs. cited by
applicant .
"International Application Serial No. PCT/US2016/047360, Written
Opinion dated Nov. 21, 2016", 8 pgs. cited by applicant .
L, Perraud, "Accumulation of Free ADP-ribose from Mitochondria
Mediates Oxidative Stress-induced Gating of TRPM2 Cation Channels",
Journal of Biological Chemistry, vol. 280, No. 7, (Feb. 18, 2005),
6138-6148. cited by applicant .
Mohammed, M Mohammed, et al., "Evaluation of the Clinical use of
Metformin or Pioglitazone in Combination with Meloxicam in Patients
with Knee Osteoarthritis; using Knee Injury and Osteoarthritis
outcome Score", Iraqi J Pharm Sci, vol. 23, No. 2 (Jan. 14, 2015),
13-26. cited by applicant .
Mustafa, Naziroglu, "New Molecular Mechanisms on the Activation of
TRPM2 Channels by Oxidative Stress and ADP-Ribose", Neurochemical
Research, Kluwer Academic Publishers-Plenum Publishers, NE vol. 32,
No. 11, (Jun. 12, 2007), 1990-2001. cited by applicant .
Todd, O McKinley, et al., "Mitochondrial Based Treatments that
Prevent Post-Traumatic Osteoarthritis in a Translational Large
Animal intraarticular Fracture Survival Model Principal
Investigator: Distribution Statement: Approved for Public Release;
Distribution Uniimited", [Online] retrieved from the Internet::
<URL:http://www.dti c.mi 1/get-tr-doc/pdf? Loc ati
on=U2&doc=GetTRDoc.pdf&AD=ADA592443>, (Jan. 28, 2014).
cited by applicant .
"International Application Serial No. PCT/US2016/047360,
Supplementary International Search Report dated Nov. 30, 2017", 37
pgs. cited by applicant .
"International Application Serial No. PCTUS2016047360 International
Preliminary Report on Patentability dated Mar. 1, 2018", 10 pgs.
cited by applicant .
Brouillette, M J, et al., "Strain-Dependent Oxidant Release in
Articular Cartilage Originates from Mitochondria", Biomech Model
Mechanobiol. 13(3), (Jun. 2014), 565-572. cited by applicant .
Brouillette, Marc James, "Static Compressive Stress Induces
Mitochondrial Oxidant Production in Articular Cartilage (Thesis)",
A thesis submitted in partial fulfillment of the requirements for
the Master of Science degree in Biomedical Engineering in the
Graduate College of the University of Iowa, (May 2012), 51 pgs.
cited by applicant .
Coleman, M, "Complex I inhibition after intra-articular fracture
prevents rapid progression of osteoarthritis in a porcine model
(Abstract with graphs)", 63rd Annual Meeting of the Orthopaedic
Research Society, San Diego, California, (2017), 1 pg. cited by
applicant .
Coleman, M, et al., "Complex I Inhibition after Intra-articular
Fracture Prevents Rapid Progression of Osteoarthritis in a Porcine
Model (Poster)", 63rd Annual Meeting of the Orthopaedic Research
Society, San Diego, California, (2017), 1 pg. cited by applicant
.
Coleman, M, "Differential Effects of Superoxide Dismutase Mimetics
after Mechanical Overload of Articular Cartilage (Abstract)",
Antioxidants, 6(4), (2017), 2 pgs. cited by applicant .
Coleman, M, et al., "Injurious Loading of Articular Cartilage
Compromises Chondrocyte Respiratory Function (Abstract)", Arthritis
Rheumatol, 68(3, (2016), 2 pgs. cited by applicant .
Coleman, M, et al., "Intraarticular Administration of
N-Acetylcysteine Alleviates Acute Oxidative Stress Following
Intraarticular Fracture (Abstract)", Oberly Symposium, Iowa City,
IA, (2015), 1 pg. cited by applicant .
Coleman, M, "Intraarticular Administration of N-Acetylcysteine and
Glycyrrhizin Alleviates Acute Oxidative Stress Following
Intraarticular Fracture (Abstract)", Orthopaedic Research Society
Annual Meeting, Las Vegas, Nevada, (2015), 1 pg. cited by applicant
.
Coleman, M, "Intraarticular Administration of N-Acetylcysteine and
Glycyrrhizin Alleviates Acute Oxidative Stress Following
Intraarticular Fracture (Poster)", Orthopaedic Research Society
Annual Meeting, Las Vegas, Nevada, (2015), 1 pg. cited by applicant
.
Coleman, M, "Intraarticular Administration of N-Acetylcysteine
Prevents Progression of Post-Traumatic Osteoarthritis in a Large
Animal Model of Intraarticular Fracture (Abstract)", Society for
Free Radical Biology and Medicine, Boston, Massachusetts, (2015), 1
pg. cited by applicant .
Coleman, M, "Intraarticular Administration of N-Acetylcysteine
Prevents Progression of Post-Traumatic Osteoarthritis in a Large
Animal Model of Intraarticular Fracture (Presentation)", Society
for Free Radical Biology and Medicine, Boston, Massachusetts,
(2015), 29 pgs. cited by applicant .
Coleman, M, et al., "N-Acetylcysteine Prevents Acute Chondrocyte
Injury and Dysfunction Associated with Osteoarthritic Progression
after Intraarticular Fracture (Poster)", Military Health System
Research Symposium, Fort Lauderdale, Florida, (2015), 1 pg. cited
by applicant .
Coleman, M, "Osteoarthritis in Porcine Intraarticular Fracture
Model Reveals Mitochondrial Features Similar to Human Disease
(Abstract)", Annual Meeting of the Orthopaedic Research Society,
Orlando, Florida, (2016), 1 pg. cited by applicant .
Coleman, M, "Osteoarthritis in Porcine Intraarticular Fracture
Model Reveals Mitochondrial Features Similar to Human Disease
(Poster)", Annual Meeting of the Orthopaedic Research Society,
Orlando, Florida, (2016), 1 pg. cited by applicant .
Coleman, M, et al., "Overloading Healthy Articular Cartilage
Induces Mitochondrial Dysfunction Reminiscent of Late Stage
Osteoarthritis (Abstract)", Orthopaedic Research Society Annual
Meeting, New Orleans, Louisiana, (2014), 1 pg. cited by applicant
.
Coleman, M, et al., "Overloading Healthy Articular Cartilage
Induces Mitochondrial Dysfunction Reminiscent of Late Stage
Osteoarthritis (Poster)", Orthopaedic Research Society Annual
Meeting, New Orleans, Louisiana, (2014), 1 pg. cited by applicant
.
Coleman, M, et al., "Targeting mitochondrial responses to
intra-articular fracture to prevent posttraumatic osteoarthritis",
Science Translational Medicine, 10, Issue 427, (Feb. 2018), 15 pgs.
cited by applicant .
Coleman, Mitchell, et al., "Complex I Inhibition after
Intra-articular Fracture Prevents Rapid Progression of
Osteoarthritis in a Porcine Model (Abstract)", OARSI, (2017), 1 pg.
cited by applicant .
Coleman, Mitchell, et al., "Differential Effects of Superoxide
Dismutase Mimetics after Mechanical Overload of Articular
Cartilage", Antioxidants 6(4), (2017), 10 pgs. cited by applicant
.
Coleman, Mitchell, et al., "Loading of Articular Cartilage
Compromises Chondrocyte Respiratory Function", Arthritis Rheumatol,
68(3), (Mar. 2016), 662-671. cited by applicant .
Coleman, Mitchell, "Mitochondrial Responses to Intraarticular
Fracture are a Disease-Modifying Target for Post-Traumatic
Osteoarthritis Prevention", Nature Medicine, (Mar. 2017). cited by
applicant .
Coleman, Mitchell, et al., "N-Acetylcysteine Prevents Acute
Chondrocyte Injury and Dysfunction Associated with Osteoarthritic
Progression after Intraarticular Fracture (Abstract)", Military
Health System Research Symposium, Fort Lauderdale, Florida, (2015),
1 pg. cited by applicant .
Coleman, Mitchell, "Three Critical Considerations for Translating
Redox Therapies: Location, Location, Location (Presentation)",
(2017), 55 pgs. cited by applicant .
Compton, Jocelyn, et al., "Sirtuin-1 Augments Chondrogenic
Progenitor Cell Activity in an Acute Cartilage Injury Model
(Poster)", ORS, (2018), 1 pg. cited by applicant .
Goetz, Jessica, et al., "Time-Dependent Loss of Mitochondrial
Function Precedes Progressive Histologic Cartilage Degeneration in
a Rabbit Meniscal Destabilization Model", J Orthop Res., 35(3),
(2017), 590-599. cited by applicant .
Goodwin, Wendy, et al., "Rotenone Prevents Impact-Induced
Chondrocyte Death", Journal of Orthopaedic Research 28(8), (2010),
1057-1063. cited by applicant .
Jubeck, Brian, et al., "Promotion of Articular Cartilage Matrix
Vesicle Mineralization by Type I Collagen", Arthritis Rheum. 58(9),
(2008), 2809-2817. cited by applicant .
Martin, James, et al., "N-Acetylcysteine Inhibits Post-Impact
Chondrocyte Death in Osteochondral Explants", Journal of Bone and
Joint Surgery, vol. 91-A, No. 8, (2009), 1890-1897. cited by
applicant .
Sauter, Ellen, et al., "Cytoskeletal Dissolution Blocks Oxidant
Release and Cell Death in Injured Cartilage", Journal of
Orthopaedic Research, 30(4), (2012), 593-598. cited by applicant
.
Sigaeva, N, et al., "Chemical modification of hyaluronic acid and
its application medicine (with machine translation)", vol. 17. No.
3. Herald of Bashkir University, (2012), 1220-1241. cited by
applicant .
Wolff, K, "Mechanical Stress and ATP Synthesis Are Coupled by
Mitochondrial Oxidants in Articular Cartilage (Abstract)", J Orthop
Res 31(2), (2013), 191-196. cited by applicant .
Wolff, Katherine, et al., "Mechanical Stress and ATP Synthesis Are
Coupled by Mitochondrial Oxidants in Articular Cartilage", Journal
of Orthopaedic Research, 31(2), (2013), 191-196. cited by applicant
.
Badwaik, V., et al., "Efficient pDNA Delivery Using Cationic
2-Hydroxypropyl-.beta.-Cyclodextrin Pluronic-Based Polyrotaxanes",
Macromol Biosci.; 16(1):63-73, (Jan. 2016), (Abstract Only). cited
by applicant .
Bahadur, A, et al., "NaCl-triggered self-assembly of hydrophilic
poloxamine block copolymers", Int J Pharm.; 494(1):453-62, (Oct.
15, 2015), (Abstract Only). cited by applicant .
Brunori, M., et al., "Nitric oxide and the respiratory enzyme",
Biochim Biophys Acta, 1757(9-10), (Sep.-Oct. 2006), (Abstract
Only). cited by applicant .
Glinka, Y., et al., "Nature of inhibition of mitochondrial
respiratory complex I by 6-Hydroxydopamine", J Neurochem. 66(5),
(May 1996), (Abstract Only). cited by applicant .
Hao, S., et al., "Mitochondrion-Targeted Peptide SS-31 Inhibited
Oxidized Low-Density Lipoproteins-Induced Foam Cell Formation
through both ROS Scavenging and Inhibition of Cholesterol Influx in
RAW264.7 Cells", Molecules.; 20(12):21287-97, (Dec. 1, 2015),
(Abstract Only). cited by applicant .
James, AD, et al., "The Plasma Membrane Calcium Pump in Pancreatic
Cancer Cells Exhibiting the Warburg Effect Relies on Glycolytic
ATP", J Biol Chem.; 290(41): 24760-71, (Oct. 2015), (Abstract
Only). cited by applicant .
Kerkhofs, S., et al., "Self-Assembly of Pluronic F127-Silica
Spherical Core-Shell Nanoparticles in Cubic Close-Packed
Structures", Chem Mater.; 27(15):5161-5169, (Aug. 11, 2015),
(Abstract Only). cited by applicant .
Moncada, PS, "Nitric Oxide and Oxygen: Actions and Interactions in
Health and Disease", Redox Biol.; 5:421, (Aug. 2015), (Abstract
Only). cited by applicant .
Muller, M., et al., "Nanostructured Pluronic hydrogels as bioinks
for 3D bioprinting", Biofabrication.; 7(3), (Aug. 2015), (Abstract
Only). cited by applicant .
Novakofski, KD, et al., "Joint-dependent response to impact and
implications for posttraumatic", Osteoarthritis Cartilage:
23(7):1130-7, (Jul. 2015), (Abstract Only). cited by applicant
.
Roma, MI, et al., "Tetronic.RTM. 904-containing polymeric micelles
overcome the overexpression of ABCG2 in the blood-brain barrier of
rats and boost the penetration of the antiretroviral efavirenz into
the CNS", Nanomedicine (Lond).; 10(15):2325-37, (2015), (Abstract
Only). cited by applicant .
Sandez-Macho, I., et al., "Interaction of poloxamine block
copolymers with lipid membranes: Role of copolymer structure and
membrane cholesterol content", Colloids Surf B Biointerfaces;
133:270-7, (Sep. 2015), (Abstract Only). cited by applicant .
Sharma, S., et al., "Investigating the role of Pluronic-g-Cationic
polyelectrolyte as functional stabilizer for nanocrystals: Impact
on Paclitaxel oral bioavailability and tumor growth", Acta
Biomater.; 26:169-83, (Oct. 2015), (Abstract Only). cited by
applicant .
Zhang, W., et al., "Involvement of ROS-mediated mitochondrial
dysfunction and SIRT3 down-regulation in
tris(2-chloroethyl)phosphate-induced cell cycle arrest", Toxicol
Res (Camb).; 5(2):461-470, (Dec. 14, 2015), (Abstract Only). cited
by applicant .
Fakhari, A, et al., "Applications and emerging trends of hyaluronic
acid in tissue engineering, as a dermal filler and in
osteoarthritis treatment (Abstract)", Acta Biomater, 9:7081,
(2013), 1 pg. cited by applicant .
Koh, Minsoo, et al., "A novel metformin derivative, HL010183,
inhibits proliferation and invasion of triple-negative breast
cancer cells (Abstract)", vol. 21, Issue 8, (2013), 2 pgs. cited by
applicant .
Sogame, Yoshihisa, "A comparison of uptake of metformin and
phenformin mediated by hOCT1 in human hepatocytes (Abstract)",
Biopharm. Drug Dispos., 30:476, (2009), 2 pgs. cited by applicant
.
"European Application Serial No. 16757452.4, Response filed Oct. 8,
2018 to Communication Pursuant to Rules 161(1) and 162 EPC dated
Mar. 29, 2018", w English Claims, 14 pgs. cited by applicant .
"Chinese Application Serial No. 201680056186.X, Voluntary Amendment
filed Oct. 8, 2018", w English Claims, 13 pgs. cited by applicant
.
"European Application Serial No. 16757452.4, Communication pursuant
to Article 94(3) EPC dated Mar. 14, 2019", 6 pgs. cited by
applicant.
|
Primary Examiner: Wax; Robert A
Assistant Examiner: Truong; Quanglong N
Attorney, Agent or Firm: Schwegman Lundberg & Woessner,
P.A.
Government Interests
STATEMENT OF GOVERNMENT RIGHTS
This invention was made with government support under grant
W81XWH-11-1-0583 awarded by the Department of Defense. The
government has certain rights in the invention.
Parent Case Text
CLAIM FOR PRIORITY
This application is a continuation of and claims the benefit of
priority to International Application No. PCT/US2016/047360, filed
Aug. 17, 2016, which claims the benefit of priority to U.S.
Provisional Patent Application Ser. No. 62/207,059, filed Aug. 19,
2015, the disclosure of which is incorporated by reference herein.
Claims
What is claimed is:
1. An injectable composition comprising a hydrogel comprising
hyaluronic acid having a molecular weight of 1 million Da or
greater and 15% wt/vol to about 20% wt/vol F127, and an amount of
amobarbital or a derivative thereof effective to inhibit
post-traumatic osteoarthritis.
2. The composition of claim 1 wherein the hyaluronic acid is
present in the composition from about 0.01% (wt/vol) and up to
about 2.0% (wt/vol).
3. The composition of claim 1 wherein the hyaluronic acid is
present at about 0.2% wt/vol to about 1.0% wt/vol.
4. The composition of claim 1 wherein the hydrogel further
comprises N-isopropyl acrylamide polymer, a polysaccharide other
than hyaluronic acid, hydroxypropylcellulose, karya gum, guar gum,
gellan gum, alginate, ethylhydroxyethylcellulose,
poly(ethyleneoxide-b-propylene oxide-b-ethylene oxide), a
PLURONICS.RTM. polymer, a poly(ethylene glycol)/poly(D,L-lactic
acid-co-glycolic acid) block co-polymer, a polyphosphazine, a
polyacrylate, a TETRONICS.TM. polymer, or a polyethylene
oxide-polypropylene glycol block copolymer.
5. The composition of claim 1 wherein the derivative comprises
pentobarbital, secobarbital, phenobarbital, or barbital.
6. The composition of claim 1 wherein the amount inhibits
mitochondrial dysfunction or chondrocyte energy dysfunction.
7. The composition of claim 1 wherein the amobarbital or a
derivative thereof scavenges mitochondrial oxidants or prevents
their formation, or stimulates glycolytic ATP production.
8. A method to inhibit chondrocyte death and improve chondrocyte
function after injury in a mammal, comprising locally administering
an effective amount of the composition of claim 1 to an injured
joint of a mammal.
9. The method of claim 8 wherein the composition is injected.
10. The method of claim 8 wherein the composition comprises
amobarbital, pentobarbital, secobarbital, or phenobarbital.
11. The method of claim 8 wherein the amount inhibits mitochondrial
dysfunction or chondrocyte energy dysfunction.
12. The method of claim 8 wherein the administration reduces ROS
production in cartilage.
13. The method of claim 8 wherein the administration is within 4
days of the injury.
14. The method of claim 8 wherein the administration is with 12
hours of the injury.
15. The method of claim 8 wherein the administration is with 5
hours of the injury.
16. The method of claim 8 wherein the injury is a knee, hip, ankle
or elbow injury.
17. A method to inhibit post-traumatic osteoarthritis in a mammal,
comprising locally administering an effective amount of the
composition of claim 1 to a mammal at risk of posttraumatic
osteoarthritis as a result of injury to cartilage.
18. The method of claim 17 wherein the composition is injected.
19. The method of claim 17 wherein the composition comprises
amobarbital, pentobarbital, secobarbital, or phenobarbital.
20. The method of claim 17 wherein the administration is within 4
days of the injury.
21. The method of claim 17 wherein the mammal has an injured
joint.
22. The method of claim 17 wherein the administration is with 12
hours of the injury.
23. The method of claim 17 wherein the administration is with 5
hours of the injury.
24. The method of claim 17 wherein the injury is a knee, hip, ankle
or elbow injury.
25. The method of claim 17 wherein the mammal is a human.
Description
BACKGROUND
The pain, immobility, and general disability associated with
osteoarthritis are familiar to most people who reach old age.
Post-traumatic osteoarthritis (PTOA) is a profoundly accelerated
form of arthritis associated with traumatic injuries to joint
articular surfaces, leading to disease progression well before
patients are considered good candidates for joint replacement
approaches common to orthopaedic medicine. Because patients are
often injured relatively young and there are presently no viable
alternatives to joint replacement, patients with PTOA often suffer
disability and morbidity comparable to chronic heart disease
patients.
Natural methods for treating PTOA include decreasing load and
stress on the injured joint or increasing comfort and
functionality. For example, weight loss, low impact exercise, and
strengthening muscles surrounding the joint may improve PTOA.
However, these approaches do not cure or prevent PTOA and may not
be fully effective.
Non-steroidal, anti-inflammatory medicines (NSAIDS) are used to
decrease pain and inflammation associated with PTOA, although
NSAIDs can cause stomach irritation and kidney, liver or heart
problems. Moreover, NSAIDs likely do not prevent PTOA.
Antioxidants, another class of compounds used to treat PTOA,
stabilize or deactivate reactive oxygen species (ROS) before they
attack cells. Nevertheless, there is skepticism about the benefit
of antioxidants and there are potentially harmful side effects if
anti-oxidants are taken in excess.
Other methods used to treat PTOA include the administration of
cortisone and hylamers which act like artificial joint fluid after
injection. However, cortisone can cause elevation of heart rate and
blood sugar and should not be given too often. In addition,
cortisone is not preventative. While corticosteroid injections are
anti-inflammatory, the potential benefit or adverse effects of that
injection for traumatic injury have not been resolved. Another
approach is the use of platelet-rich plasma injections.
Injection of a patient's own platelets leads to release of growth
factors and attraction of regenerative cells to the site of injury.
This type of injection is not preventative and does not work for
all PTOA patients. Moreover, details on dosage, frequency of
injection, and other important parameters have yet to be worked out
for platelet rich plasma administration. A further type of
injection is an amniotic membrane stem cell injection. While this
injection is anti-inflammatory, thus providing pain relief, and
results in replacement of damaged cells due to release of growth
factors, it is not preventative and does not target ROS.
If non-surgical methods are ineffective, surgical methods may be
employed to restore the joint after PTOA. The surgery may include
cleaning out, reconstructing or replacing the worn out joint
surfaces. As with other surgeries, there can be surgical
complications, e.g., infection and damage to surrounding
structures, blood clots, heart attack, and stroke, and the eventual
wearing out or loosening of implants.
SUMMARY
The present disclosure provides an injectable composite hydrogel
comprising a polysaccharide, e.g., a natural polysaccharide such as
hyaluronic acid, hydroxypropylcellulose, karya gum (KG), guar gum
(GUG), or gellan gum (GEG), a semi-synthetic polysaccharide or a
synthetic polysaccharide, and a synthetic polymer, e.g., F127,
whose reverse-thermal properties cause the composite to become firm
once injected (preventing leakage from the site of injection such
as a joint), and a compound useful to prevent, inhibit or treat
PTOA. In one embodiment, the compound reversibly inhibits the
respiratory enzyme complex I, a key mediator of chondrocyte injury
after impact. In one embodiment, the hydrogel comprises an
effective amount of amobarbital, e.g., from about 0.25 mM to about
50 mM or about 1.25 mM to about 10 mM, metformin
(N,N,-dimethylbiguanide) a biguanide derivative,
N,N-diethylbiguanide, N,N,-dipropylbiguanide, phenformin (Sogame et
al., Biopharm. Drug Dispos., 30:476 (2009)), or HL010183 (Koh et
al., Bioorg. Med. Chem., 21:2305 (2013)), or adenosine diphosphate
ribose or a derivative thereof. In one embodiment, the volume
administered is about 0.1 mL to about 15 mL, e.g., about 1 mL to
about 10 mL or about 2 mL to about 5 mL. The combination of
materials in the hydrogel offers a practical advantage, for
instance, in enabling health care providers to protect articular
tissue acutely after injury. Also, the use of compounds that
reversibly inhibit the respiratory enzyme complex I to alter
articular cartilage provides for chondroprotection after injury and
eventual reestablishment of normal activity of the respiratory
enzyme complex I.
The disclosure provides an injectable composition comprising a
composite reverse-temperature sensitive hydrogel comprising a
biopolymer, such as a polysaccharide, and a synthetic polymer, and
a compound in an amount that optionally reversibly inhibits
respiratory enzyme complex I. In one embodiment, the hydrogel
includes about 0.2 wt/vol to about 4% wt/vol HA In one embodiment,
the polysaccharide comprises hyaluronic acid. In one embodiment,
the synthetic polymer comprises a poloxamer, e.g., F127. In one
embodiment, the hydrogel includes about 15% wt/vol to about 20%
wt/vol F127. In one embodiment, the compound comprises amobarbital.
In one embodiment, the amount of the compound in the hydrogel
inhibits mitochondrial dysfunction or chondrocyte energy
dysfunction. In one embodiment, the compound scavenges
mitochondrial oxidants or prevents their formation, or stimulates
glycolytic ATP production In one embodiment, the hydrogel comprises
N-isopropyl acrylamide polymer, ethylhydroxyethylcellulose,
poly(etheylene oxide-b-propylene oxide-b-ethylene oxide),
poloxamers, PLURONICS.RTM. polymers, poly(ethylene
glycol)/poly(D,L-lactic acid-co-glycolic acid) block co-polymers,
polysaccharides, alginate, polyphosphazines, polyacrylates,
TETRONICS.TM. polymers, or polyethylene oxide-polypropylene glycol
block copolymers. In one embodiment, the polysaccharride comprises
hyaluronic acid of about or greater than 1.5 M Dalton. In one
embodiment, the MW is about 1,600,000 to 3,200,000, or about
1,900,000 to 3,900,000.
In one embodiment, the polysaccharide comprises
hydroxypropylcellulose, karya gum (KG), guar gum (GUG), or gellan
gum (GEG). In one embodiment, the polysaccharide is present in the
hydrogel at about 0.2% (wt/vol) to about 1.0% (wt/vol).
In one embodiment, the composition is a reverse
temperature-sensitive hydrogel (one that is non-viscous at "low"
temperature, e.g., at or below room temperature, e.g., about
70.degree. F. or less. The low initial viscosity allows the
hydrogel to coat all the cartilage surfaces through the joint
before it sets (i.e., the viscosity increases at temperatures above
room temperature, e.g., about 80.degree. F. or greater including
human body temperature such as about 98.degree. F.), which provides
for superior retention in the joint and substantially improves the
bioavailablity of the compound dissolved in the gel. Reverse
temperature-sensitive hydrogels, which have initial viscosities of
about 100 to about 160 or about 80 to about 200, e.g., about 120 to
about 140, Pascal Seconds, may be administered using a 22 to 24
gauge needle, e.g., a 22 gauge needle. In contrast, non-reverse
temperature-sensitive hydrogels require large bore needles and do
not evenly distribute in the joint due to their high initial
viscosity.
Also provided is a method to prevent or inhibit chondrocyte death
and improve chondrocyte function after injury in a mammal. The
method includes administering an effective amount of the
composition to a mammal having the injury. Further provided is a
method to prevent or inhibit post-traumatic osteoarthritis in a
mammal. The method includes administering an effective amount of
the composition a mammal at risk of posttraumatic osteoarthritis.
In one embodiment, the composition comprises hyaluronic acid. In
one embodiment, the composition comprises F127. In one embodiment,
the composition comprises amobarbital. In one embodiment, the
amount administered inhibits mitochondrial dysfunction or
chondrocyte energy dysfunction. In one embodiment, the compound
administered scavenges mitochondrial oxidants or prevent their
formation, in addition to stimulating glycolytic ATP production. In
one embodiment, the administration is within 1, 2, 3, 4 or 5 days
of the injury. In one embodiment, the mammal has an injured joint.
In one embodiment, the administration is with 1, 2, 3, 4, 5, 6, 7,
8, 9, 10 11, or 12 hours of the injury.
DETAILED DESCRIPTION OF THE FIGURES
FIG. 1 is a schematic of the electron transport chain. Electrons
from donor molecules are transferred through protein complexes. As
electrons are transferred, hydrogen ions are pumped across the
inner membrane of the mitochondria, and as the hydrogen atoms fall
back over the inner membrane, they generate ATP.
FIG. 2 is a schematic of reactive oxygen species production.
FIG. 3 is a schematic of steps in the progression to post-traumatic
osteoarthritis.
FIG. 4 shows complex I activity in the presence or absence of
amobarbital.
DETAILED DESCRIPTION
Definitions
"Hydrogel" as used herein means a water insoluble, naturally or
chemically-induced cross-linked, three-dimensional network of
polymer chains plus water that fills the voids between polymer.
Cartilage, Electron Transport and PTOA
Articular cartilage is the smooth, white tissue that covers the
ends of bones where they come together to form joints. It allows
the bones to glide over each other with very little friction, and
acts as a cushion. Injured, inflamed, or damaged cartilage does not
heal itself well due to lack of a blood supply, resulting in
symptoms such as pain and limited movement leading to joint damage
and deformity. Chondrocytes are cells found in cartilage connective
tissue they produce and maintain the cartilage matrix. Under normal
circumstances, cartilage wears down over time and chondrocytes
replace and repair it as needed.
Chondrocyes, like all cells, contain mitochondria. Mitochondria
generate ATP through the Electron Transport Chain (ETC) (FIG. 1).
Sometimes, harmful substances called reactive oxygen species (ROS)
(FIG. 2) are created through the ATP generation process and are
formed by Respiratory Complex I in the Electron Transport Chain.
ROS can act as signaling molecules and signal healthy chondrocytes
to undergo apoptosis (cell suicide), depending on the severity and
length of exposure, which leads to osteoarthritis.
Osteoarthritis is wearing out of joint surface cartilage over time.
Post-traumatic osteoarthritis (PTOA) is wearing out of a joint that
has had any kind of physical injury. PTOA is a debilitating
consequence of intraarticular fractures. Patient outcomes after
intraarticular fractures have not improved significantly in spite
of improved surgical techniques. PTOA is relatively common: As of
2006 approximately 12% of the overall prevalence of symptomatic OA
was attributable to PTOA of the hip, knee, or ankle. This
corresponds to approximately 5.6 million individuals in the United
States being affected by PTOA. The corresponding aggregate
financial burden specifically of PTOA is $3.06 billion annually, or
approximately 0.15% of the total U.S. health care direct cost
outlay.
Compositions and Methods to Prevent, Inhibit or Treat PTOA
Inhibiting electron transport and associated oxidant production by
chondrocytes after impact injuries associated with PTOA prevents
cell death and dysfunction. Accordingly, by muting chondrocyte
mitochondrial metabolism acutely after traumatic injury using
compounds that inhibit respiratory enzyme complex I (and also
decrease ROS) that are delivered intra-articularly in a hydrogel
vehicle, the treatment is confined to the joint capsule and
prevents leaking out of any joint disruptions present. This allows
controlled local delivery of an effective pharmaceutical in a
manner that minimizes exposure to the rest of the body. For
example, amobarbital is a barbiturate derivative used to produce
relaxation, sleep, anesthesia, and anticonvulsant effects. It
inhibits respiratory complex I, leading to a decrease in ROS.
Because the effect of amobarbital in inhibiting mitochondrial
electron transport is reversible, unlike rotenone or other more
toxic alternatives, transient manipulation of chondrocyte
metabolism in this manner can prevent chondrocyte injury and death,
as well as subsequent disease, while avoiding toxic insult to the
patient due to return of oxidative metabolism.
The present compositions and method are useful to prevent, inhibit
or treat PTOA, and may substantially lower or eliminate treatment
costs and morbidities associated with other more invasive
approaches that require multiple surgical procedures and/or cell
harvests.
Hydrogels and Polymers Useful in Hydrogels
Hydrogels can be classified as those with crosslinked networks
having permanent junctions or those with physical networks having
transient junctions arising from polymer chain entanglements or
physical interactions, e.g., ionic interactions, hydrogen bonds or
hydrophobic interactions. Natural materials useful in hydrogels
include natural polymers, which are biocompatible, biodegradable,
support cellular activities, and may include proteins like fibrin,
collagen or gelatin, and/or polysaccharides like hyaluronic acid,
starch, alginate or agarose. Synthetic polymers useful in hydrogels
are prepared by chemical polymerization and include by way of
example poloxamers, acrylic acid, hydroxyethyl-methacrylate (HEMA),
vinyl acetate, and methacrylic acid (MAA).
Various methods may be used to prepare hydrogels, e.g.,
crosslinkers, copolymerization of monomers using multifunctional
co-monomer, cross linking of linear polymers by irradiation or by
chemical compounds. Monomers contain an ionizable group that can be
ionized or can undergo a substitution reaction after the
polymerization is completed. Exemplary crosslinkers are
glutaraldehyde, calcium chloride and oxidized konjac glucomannan
(DAK).
Some classes of hydrogels include (a) homopolymeric hydrogels which
are derived from a single species of monomer. Homopolymers may have
cross-linked skeletal structure depending on the nature of the
monomer and polymerization technique; (b) copolymeric hydrogels
which are comprised of two or more different monomer species with
at least one hydrophilic component, arranged in a random, block or
alternating configuration along the chain of the polymer network;
(c) multipolymer interpenetrating polymeric hydrogel (IPN) which is
made of two independent cross-linked synthetic and/or natural
polymer components, contained in a network form. In semi-IPN
hydrogel, one component is a cross-linked polymer and other
component is a non-cross-linked polymer.
Biodegradable hydrogels as a delivery vehicle have the advantage of
being environmentally friendly to the human body (due to their
biodegradability) and of providing more predictable, controlled
release of the impregnated drugs. Hydrogels are of special interest
in biological environments since they have a high water content as
is found in body tissue and are highly biocompatible. Hydrogels and
natural biological gels have hydrodynamic properties similar to
that of cells and tissues. Hydrogels minimize mechanical and
frictional irritation to the surrounding tissue because of their
soft and compliant nature. Therefore, hydrogels provide a far more
user-friendly delivery vehicle than the relatively hydrophobic
carriers like silicone, or vinyl acetate.
Biocompatible materials that may be present in a hydrogel include,
e.g., permeable configurations or morphologies, such as polyvinyl
alcohol, polyvinylpyrrolidone and polyacrylamide, polyethylene
oxide, poly(2-hydroxyethyl methacrylate); natural polymers such as
polysaccharides, gums and starches; and include
poly[.alpha.(4-aminobutyl)]-1-glycolic acid, polyethylene oxide,
polyorthoesters, silk-elastin-like polymers, alginate, EVAc
(poly(ethylene-co-vinyl acetate), microspheres such as poly (D,L
-lactide-co-glycolide) copolymer and poly (L -lactide),
poly(N-isopropylacrylamide)-b-poly(D,L -lactide), a soy matrix such
as one cross-linked with glyoxal and reinforced with a bioactive
filler, e.g., hydroxylapatite,
poly(epsilon-caprolactone)-poly(ethylene glycol) copolymers,
poly(acryloyl hydroxyethyl) starch, polylysine-polyethylene glycol,
or agarose.
In one embodiment, the hydrogel includes poloxamers,
polyacrylamide, poly(2-hydroxyethyl methacrylate),
carboxyvinyl-polymers (e.g., Carbopol 934, Goodrich Chemical Co.),
cellulose derivatives, e.g., methylcellulose, cellulose acetate and
hydroxypropyl cellulose, polyvinyl pyrrolidone or polyvinyl
alcohols, or combinations thereof.
In some embodiments, the hydrogel includes collagen, e.g.,
hydroxylated collagen, fibrin, polylactic-polyglycolic acid, or a
polyanhydride. Other examples include, without limitation, any
biocompatible polymer, whether hydrophilic, hydrophobic, or
amphiphilic, such as ethylene vinyl acetate copolymer (EVA),
polymethyl methacrylate, polyamides, polycarbonates, polyesters,
polyethylene, polypropylenes, polystyrenes, polyvinyl chloride,
polytetrafluoroethylene, N-isopropylacrylamide copolymers,
poly(ethylene oxide)/poly(propylene oxide) block copolymers,
poly(ethylene glycol)/poly(D,L-lactide-co-glycolide) block
copolymers, polyglycolide, polylactides (PLLA or PDLA),
poly(caprolactone) (PCL), or poly(dioxanone) (PPS).
In another embodiment, the biocompatible material includes
polyethyleneterephalate, polytetrafluoroethylene, copolymer of
polyethylene oxide and polypropylene oxide, a combination of
polyglycolic acid and polyhydroxyalkanoate, gelatin, alginate,
poly-3-hydroxybutyrate, poly-4-hydroxybutyrate, and
polyhydroxyoctanoate, and polyacrylonitrilepolyvinylchlorides.
In one embodiment, the following polymers may be employed, e.g.,
natural polymers such as alginate, agarose, starch, fibrin,
collagen, gelatin, chitin, glycosaminoglycans, e.g., hyaluronic
acid, dermatan sulfate and chrondrotin sulfate, and microbial
polyesters, e.g., hydroxyalkanoates such as hydroxyvalerate and
hydroxybutyrate copolymers, and synthetic polymers, e.g.,
poly(orthoesters) and polyanhydrides, and including homo and
copolymers of glycolide and lactides (e.g., poly(L-lactide,
poly(L-lactide-co-D,L-lactide), poly(L-lactide-co-glycolide,
polyglycolide and poly(D,L-lactide), pol(D,L-lactide-coglycolide),
poly(lactic acid colysine) and polycaprolactone.
In one embodiment, the hydrogel comprises a poloxamer
(polyoxyethylene, polyoxypropylene block copolymers, e.g.,
poloxamer 127, 231, 182 or 184).
Exemplary Components for Use in Hydrogels to Prevent, Inhibit or
Treat PTOA
In one embodiment, the hydrogels useful in the compositions and
methods of the invention are synthesized from a naturally occurring
biodegradable, biocompatible, and hydrophilic polysaccharide, and a
synthetic biocompatible polymer, such as poloxamers, polylactide
("PLA") polyglycolide ("PGA"), or poly(lactic acid co-glycolic
acid) ("PLGA").
The composition of the invention that forms a hydrogel, e.g., a
reverse temperature-sensitive hydrogel, includes a polysaccharide,
including chemically cross linked polysaccharides and a synthetic
or natural polymer, and a compound that reversibly inhibits complex
I. One exemplary polysaccharide is hyaluronic acid (HA), a
naturally occurring co-polymer composed of the sugars glucuronic
acid and N-acetylglucosamine. Specifically, HA, also named
hyaluronan, is a high molecular weight (10.sup.5-10.sup.7 Da)
naturally occurring biodegradable polymer that is an unbranched
non-sulfated glycosaminoglycan (GAG) composed of repeating
disaccharides (.beta.-1,4-D-glucuronic acid (known as uronic acid)
and .beta.-1,3-N-acetyl-D-glucosamide). HA has an average MW of 4-5
million Da. HA can include several thousand sugar molecules in the
backbone. HA is a polyanion that can self-associate and that can
also bind to water molecules (when not bound to other molecules)
giving it a stiff, viscous quality similar to gelatin. Hylans are
cross-linked hyaluronan chains in which the carboxylic and N-acetyl
groups are unaffected. The MW of hylan A is about 6 million Da.
Hylans can be water-insoluble as a gel (e.g., hylan B).
HA's characteristics include its consistency, biocompatibility,
hydrophilicity, viscoelasticity and limited immunogenicity. The
hyaluronic acid backbone is stiffened in physiological solution via
a combination of internal hydrogen bonds, interactions with
solvents, and the chemical structure of the disaccharide. At very
low concentrations, HA chains entangle each other, leading to a
mild viscosity (molecular weight dependent). On the other hand, HA
solutions at higher concentrations have a higher than expected
viscosity due to greater HA chain entanglement that is
shear-dependent. Thus, solutions containing HA are viscous, but the
viscosity is tunable by varying HA concentration and the amount of
crosslinking. In addition to the unique viscosity of HA, the
viscoelasticity of HA is another characteristic resulting from
entanglement and self-association of HA random coils in solution.
Viscoelasticity of HA can be tied to molecular interactions which
are also dependent on concentration and molecular weight.
Exemplary HA solutions for injection are shown in Table 1, and
include include Synvisc.RTM. (high molecular weight HA due to
crosslinking), Hyalgan.RTM. (sodium hyaluronate solution), and
Orthovisc.RTM. (one of the viscosupplements with the highest HA
concentration, which has lower viscosity than Synvisc.RTM.) (the
properties of those are shown in Table 2).
TABLE-US-00001 TABLE 1 Molecular weight Brand name (Generic name)
(kDa) Durolane .RTM. (Hyaluronic acid, 2%) 1000 Fermathron .RTM.
(Sodium hyaluronate, 1%) 1000 Hyalgan .RTM. (Sodium hyaluronate,
1%) 500-730 NeoVisc .RTM. (Sodium hyaluronate, 1%) 1000 Orthovisc
.RTM. (Sodium hyaluronate, 1%) 1000-2900 Ostenil .RTM. (Sodium
hyaluronate, 1%) 1000-2000 Supartz .RTM. (Sodium hyaluronate, 1%)
620-1170 Suplasyn .RTM. (Sodium hyaluronate, 1%) 500-730 Synvisc
.RTM. (Hylan G-F 20; Crosslinked HA) 6000-7000
TABLE-US-00002 TABLE 2 Viscoelastic properties Molecular Elastic
Viscous weight modulus (G') modulus (G'') Brand name (kDa) (Pa) at
2.5 Hz (Pa) at 2.5 Hz Hyalgan .RTM. (Uncrosslinked) 500-730 0.6 3
Orthovisc .RTM. 1000-2900 60 46 (Uncrosslinked) Synvisc .RTM.
(Crosslinked 6000-7000 111 .+-. 13 25 .+-. 2 polymer)
Dextran is another polysaccharide and is formed primarily of
1,6-.alpha.-D-glucopyranosyl residues and has three hydroxyl groups
per glucose residue that could provide greater flexibility in the
formulation of hydrogels. Dextran has been widely used for many
biomedical purposes, such as plasma expander and controlled drug
delivery vehicle, because of its highly hydrophilic nature and
biocompatibility. It is also possible to incorporate dextranase in
order to facilitate biodegradation of dextran for the meeting of
specific clinical needs.
In one embodiment, the hydrogel comprises a poloxamer. Poloxamers
are nonionic triblock copolymers composed of a central hydrophobic
chain of polyoxypropylene (poly(propylene oxide)) flanked by two
hydrophilic chains of polyoxyethylene (poly(ethylene oxide))
(.alpha.-Hydro-.omega.-hydroxypoly (oxyethylene).sub.a poly
(ocypropylene).sub.b poly (olxyethylene).sub.a block copolymer,
with two hydrophilic chains of ethylene oxide chains (PEO) that
sandwich one hydrophobic propylene oxide chain (PPO) giving a
chemical formula
HO(C.sub.2H.sub.4O).sub.a(C.sub.3H.sub.6O).sub.b(C.sub.2H.sub.4O).sub.a).
For example, poloxamer 407 is a triblock copolymer consisting of a
central hydrophobic block of polypropylene glycol flanked by two
hydrophilic blocks of polyethylene glycol. Exemplary poloxamers
include but are not limited to polyethylene-propylene glycol
copolymer, e.g., Supronic, Pluronic or Tetronic a non-ionic
triblock copolymer.
The common representation of Poloxamer is indicated as `P`
succeeded by three digits where the first two digits are to be
multiplied by 100 and that gives the molecular mass of the
hydrophobic propylene oxide and the last digit is to be multiplied
by ten that gives the content of hydrophilic ethylene oxide in
percentage. Poloxamers usually have an efficient thermoreversible
property with characteristics sol-gel transition temperature. Below
the transition temperature it is present as a solution and above
this temperature the solution results in interaction of the
copolymer segment which leads to gelation. Poloxamers are non-toxic
and non-irritant.
TABLE-US-00003 TABLE 3 Ethylene Average oxide Propylene molecular
Weight % of units oxide units mass Oxyethylene Physical (n).sup.a
(n).sup.a PhEur 2005; PhEur USPNF Poloxamer Pluronic form (a) (b)
USPNF 23 2005 23 124 L 44 Liquid 10-15 18-23 2090-2360 44.8-48.6
46.7 .+-. 1.9 188 F 68 Solid 75-85 25-40 7680-9510 79.9-83.7 81.8
.+-. 1.9 237 F 87 Solid 60-68 35-40 6840-8830 70.5-74.3 72.4 .+-.
1.9 338 F108 Solid 137-146 42-47 12700-17400 81.4-84.9 83.1 .+-.
1.7 407 F127 Solid 95-105 54-60 9840-14600 71.5-74.9 73.2 .+-.
1.7
Compounds that reversibly inhibit complex I include but are not
limited to amobarbital or derivatives thereof, metformin or
derivatives thereof, or adenosine diphosphate ribose analogs that
disrupt NADH binding. However, non-reversible inhibitors of complex
I, e.g., Rotenone, Piericidin A or Rolliniastatin 1 and 2, in low
doses, may also have some benefit to cartilage after injury as a
result of altering ROS.
Formulations and Dosages
The components of the composition of the invention can be
formulated as pharmaceutical compositions and administered to a
mammalian host, such as a human patient in a variety of forms
adapted to the chosen route of administration. In one embodiment,
the components of the composition are locally administered to a
site of cartilage damage or suspected cartilage damage, or is
administered prophylactically.
In one embodiment, the components of the composition may be
administered by infusion or injection. Solutions may be prepared in
water, optionally mixed with a nontoxic surfactant. Dispersions may
also be prepared in glycerol, liquid polyethylene glycols,
triacetin, and mixtures thereof and in oils. Under ordinary
conditions of storage and use, these preparations contain a
preservative to prevent the growth of microorganisms.
The pharmaceutical dosage forms suitable for injection or infusion
may include sterile aqueous solutions or dispersions or sterile
powders comprising the active ingredient which are adapted for the
extemporaneous preparation of sterile injectable or infusible
solutions or dispersions. In all cases, the ultimate dosage form
should be sterile, fluid and stable under the conditions of
manufacture and storage. The liquid carrier or vehicle may be a
solvent or liquid dispersion medium comprising, for example, water,
ethanol, a polyol (for example, glycerol, propylene glycol, liquid
polyethylene glycols, and the like), vegetable oils, nontoxic
glyceryl esters, and suitable mixtures thereof. The prevention of
the action of microorganisms can be brought about by various
antibacterial and antifungal agents, for example, parabens,
chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In
many cases, it may be preferable to include isotonic agents, for
example, sugars, buffers or sodium chloride.
Sterile injectable solutions may be prepared by incorporating the
active agent in the required amount in the appropriate solvent with
various other ingredients, as required, optionally followed by
filter sterilization. In the case of sterile powders for the
preparation of sterile injectable solutions, the methods of
preparation include vacuum drying and the freeze drying techniques,
which yield a powder of the active ingredient plus any additional
desired ingredient present in the previously sterile-filtered
solutions.
Useful solid carriers may include finely divided solids such as
talc, clay, microcrystalline cellulose, silica, alumina and the
like. Useful liquid carriers include water, alcohols or glycols or
water-alcohol/glycol blends, in which the present compounds can be
dissolved or dispersed at effective levels, optionally with the aid
of non-toxic surfactants. Adjuvants such as antimicrobial agents
can be added to optimize the properties for a given use. Thickeners
such as synthetic polymers, fatty acids, fatty acid salts and
esters, fatty alcohols, modified celluloses or modified mineral
materials can also be employed with liquid carriers to form
spreadable pastes, gels, ointments, soaps, and the like, for
application directly to the skin of the user.
Useful dosages of the compound(s) in the composition can be
determined by comparing their in vitro activity and in vivo
activity in animal models thereof. Methods for the extrapolation of
effective dosages in mice, and other animals, to humans are known
to the art; for example, see U.S. Pat. No. 4,938,949.
Generally, the concentration of the compound(s) in a liquid
composition, may be from about 0.1-25 wt-%, e.g., from about 0.5-10
wt-%. The concentration in a semi-solid or solid composition such
as a gel or a powder may be about 0.1-5 wt-%, e.g., about 0.5-2.5
wt-%.
The amount of the compound for use alone or with other agents may
vary with the type of hydrogel, route of administration, the nature
of the condition being treated and the age and condition of the
patient, and will be ultimately at the discretion of the attendant
physician or clinician.
The components of the composition may be conveniently administered
in unit dosage form; for example, containing 5 to 1000 mg,
conveniently 10 to 750 mg, or conveniently 50 to 500 mg of active
ingredient per unit dosage form.
In general, however, a suitable dose may be in the range of from
about 0.5 to about 100 mg/kg, e.g., from about 10 to about 75 mg/kg
of body weight per day, such as 3 to about 50 mg per kilogram body
weight of the recipient per day, for example in the range of 6 to
90 mg/kg/day, e.g., in the range of 15 to 60 mg/kg/day.
The invention will be described by the following non-limiting
example.
EXAMPLE
In one embodiment, an injectable temperature-sensitive composite
hydrogel is employed where the hydrogel is liquid during injection,
then gelates when inside the body (gelates at human body
temperatures). In one embodiment, the injectable
temperature-sensitive composite hydrogel (e.g., one having
hyaluronic acid, such as Gel One which is chemically cross-linked
and has a high molecular weight, and F127) is employed to deliver a
therapeutic, for instance, the hydrogel is loaded with amobarbital
which reversibly inhibits the respiratory enzyme Complex I, a key
mediator of chondrocyte injury after impact. The hydrogel becomes
firm once injected (preventing leakage from the joint) allowing the
therapeutic to be retained in the joint region, for example, for
about 3 days after injection into the site of articular injury. In
one embodiment, the hydrogel comprises 17% (w/v) F-127 and 0.2%
(w/v) HA, and is loaded with 2.5 mM amobarbital. The
temperature-sensitive hydrogel fixes the amobarbital, which
prevents chondrocyte death, at the site of injury.
FIG. 4 shows that amobarbital directly inhibits the biochemical
activity of complex I of the electron transport chain in
chondrocytes.
REFERENCES
Martin et al., Journal of Bone and Joint Surgery, 91A:1890. Goodwin
et al., Journal of Orthopaedic Research, 28(8):1057. Wolff et al.,
Journal of Orthopaedic Research, 31:191 (2015). Sauter et al.,
Journal of Orthopaedic Research, 30:593. Jubeck, Arthritis Rheum.,
58(9):2809.
All publications, patents and patent applications are incorporated
herein by reference. While in the foregoing specification, this
invention has been described in relation to certain preferred
embodiments thereof, and many details have been set forth for
purposes of illustration, it will be apparent to those skilled in
the art that the invention is susceptible to additional embodiments
and that certain of the details herein may be varied considerably
without departing from the basic principles of the invention.
* * * * *
References